The goal of this project is to develop new or improved methods for the determination of moisture in biological products. A low residual moisture in necessary for the stability, viability and potency of the freeze-dried biological product. The residual moisture of freeze-dried biological products was first determined by the gravimetric or loss-on-drying method utilizing phosphorus pentoxide and vacuum at room temperature. This method has been optimized. For samples with uncomplicated thermogravimetric analysis (TG) curves, TG results have been shown to correlate with coulometric Karl Fischer results. Karl Fischer and TG moisture results may be different from the gravimetric moisture result for the same freeze-dried product due to the fact that different types of moisture (physically adsorbed or chemically bound moisture) are being measured. The thermogravimetric method has been used to determine the moisture content of Group A and Group C Meningococcal Polysaccharide bulks at levels of 5% to 25% moisture. Thermogravimetric/mass spectrometry (TG/MS) identified the TG transition corresponding to the loss of residual moisture in vaccines that have complex TG curves. Thermogravimetry provides precise heating conditions and weight lose information at specified temperatures, while mass spectrometry identifies volatile compounds evolved during the weight loss process. A now TG/MS interface applicable to this analysis has been developed in our laboratory. The glass tubing interface connects the quartz combustion tubs of the TG to the jet separator of the mans spectrometer. This interface allows continuous monitoring of the ion intensities of mans peaks m/e=18 (water) and m/e=44 (carbon dioxide) for the determination of residual moisture in freeze-dried biological products. Data has been collected clarifying thermograms for both Giant Short Ragweed Allergenic Extracts an well an Limulus Amebocyte Lysate, Haemophilus b Polysaccharide Conjugate Vaccines and other products.